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           Search results for: PLASMINOGEN MONOCLONAL ANTIBODIES Anti Human Plasminogen, Clone 4G6-B11   

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Monoclonal antibody to cytokeratin VKIALEVEIATY sequence motif reduces plasminogen activation in breast tumour cells.

Cytokeratins (CKs) are the main structural proteins of epithelial cells. Although they mainly form cytoplasmic structures, they are also localized at the plasma membrane or secreted from the cells. Some CKs are over-expressed in tumour cells and are used as diagnostic and prognostic biomarkers. A stable hybridoma cell line producing anti-cytokeratin monoclonal antibody (anti-CK MAb) was prepared after immunizing mice with breast cancer MCF-7 cell lysate. As shown by 2D electrophoresis, immunoblotting and mass spectroscopy, the monoclonal antibody recognizes an epitope on CK1, CK2, CK8, CK10 and CK18 in MCF-7 cells. To identify the binding site of the antibody three peptides of 12 amino acids were synthesized, each overlapping a 27 amino acid consensus sequence of the recognized CKs. Anti-CK MAb expressed high affinity for a dodecapeptide with the sequence VKIALEVEIATY, localized in the CK alpha-helical B2 domain, as shown by ELISA and surface plasmon resonance. Treatment of MCF-7 cells by anti-CK MAb impaired plasminogen activation and consequently invasiveness of the cells. Our results show that, besides their use in diagnosis, anti-cytokeratin antibodies could be used in therapy of invasive breast cancer.

2956 related Products with: Monoclonal antibody to cytokeratin VKIALEVEIATY sequence motif reduces plasminogen activation in breast tumour cells.

Anti C Reactive Protein A Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon anti HSV (II) gB IgG1 (mo anti HCMV IE pp65 IgG1 (m anti HCMV gB IgG1 (monocl HIV1 integrase antibody, Shiga Toxin 1 antibody, M Shiga Toxin 2 antibody, M Cholera toxin antibody, M

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Murine monoclonal antibodies against murine uPA receptor produced in gene-deficient mice: inhibitory effects on receptor-mediated uPA activity in vitro and in vivo.

Binding of urokinase plasminogen activator (uPA) to its cellular receptor, uPAR, potentiates plasminogen activation and localizes it to the cell surface. Focal plasminogen activation is involved in both normal and pathological tissue remodeling processes including cancer invasion. The interaction between uPA and uPAR therefore represents a potential target for anti-invasive cancer therapy. Inhibitors of the human uPA-uPAR interaction have no effect in the murine system. To enable in-vivo studies in murine cancer models we have now generated murine monoclonal antibodies (mAbs) against murine uPAR (muPAR) by immunizing uPAR-deficient mice with recombinant muPAR and screened for antibodies, which inhibit the muPA-muPAR interaction. Two of the twelve mAbs obtained, mR1 and mR2, interfered with the interaction between muPAR and the amino-terminal fragment of muPA (mATF) when analyzed by surface plasmon resonance. The epitope for mR1 is located on domain I of muPAR, while that of mR2 is on domains (II-III). In cell binding experiments using radiolabelled mATF, the maximal inhibition obtained with mR1 was 85% while that obtained with mR2 was 50%. The IC(50) value for mR1 was 0.67 nM compared to 0.14 nM for mATF. In an assay based on modified anthrax toxins, requiring cell-bound muPA activity for its cytotoxity, an approximately 50% rescue of the cells could be obtained by addition of mR1. Importantly, in-vivo efficacy of mR1 was demonstrated by the ability of mR1 to rescue mice treated with a lethal dose of uPA-activatable anthrax toxins.

2430 related Products with: Murine monoclonal antibodies against murine uPA receptor produced in gene-deficient mice: inhibitory effects on receptor-mediated uPA activity in vitro and in vivo.

Human integrin aVb3, affi IGF-1R Signaling Phospho- Insulin Receptor Phospho- T-Cell Receptor Signaling Goat Anti-Human Bradykini Goat Anti-Human, Mouse Ca Goat Anti-Rat CCKA Recept Goat Anti- Dopamine recep Goat Anti- EP1 receptor P Goat Anti-Human EP4 prost Goat Anti-Human Farnesoid Goat Anti- Glucogon recep

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Identification and characterization of a novel angiostatin-binding protein by the display cloning method.

Angiostatin is a potent anti-angiogenic protein. To examine the angiostatin-interacting proteins, we used the display-cloning method with a T7 phage library presenting human cDNAs. The specific T7 phage clone that bound to the immobilized angiostatin was isolated, and a novel gene encoding the displayed polypeptide on the isolated T7 phage was identified. The displayed angiostatin-binding sequence was expressed in E. coli as a soluble protein and purified to homogeneity. This novel angiostatin-binding region interacted specifically to angiostatin with a dissociation constant of 3.4 x 10(-7) M. A sequence analysis showed that the identified sequence was a part of the large ORF of 1,998 amino acids, whose function has not yet been characterized. A Northern analysis indicated that the gene containing the angiostatin-binding sequence was expressed differentially in the developmental stages or cell types.

1219 related Products with: Identification and characterization of a novel angiostatin-binding protein by the display cloning method.

Rabbit Anti-Rat Androgen Acyl CoA binding Protein Mouse Anti-Human Retinol S100 alpha - Rabbit polyc Human S100 Calcium Bindin ribosome binding protein ribosome binding protein calcium binding protein P SH3 domain-binding protei Guanylate-binding protein amyloid beta precursor pr Recombinant Human Angiost

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Production and characterization of monoclonal antibodies directed to the kringle V and protease domains of human apolipoprotein(a).

Production and use of anti-apolipoprotein(a) monoclonal antibodies (MAbs) specific to single copy regions in the polymorphic lipoprotein(a) (Lp(a)) has been emphasized to be important for the standardization of measurements of the coronary heart disease risk factor, Lp(a). Here, mouse MAbs were prepared against the kringle V (V) and protease (P) domains of human apolipoprotein(a) (apo(a)), which domains are present in single copy in the apo(a) molecule. The cDNA for apo(a)VP was cloned from human liver cDNA library, and the V-P recombinant protein overexpressed in Escherichia coli was used as an antigen for the antibody production. Two antibodies named as MAb(a)20 and MAb(a)23 were finally produced, and they were characterized for their binding specificity and epitopes. The specificity of the antibodies was confirmed by an immunoblotting procedure and an enzyme-linked immunoassay (ELISA). It was shown that the antibodies had little, if any, cross-reactivity with human plasminogen, which is relatively abundant in human serum and is highly homologous (85%) with apo(a) in amino acid (aa) sequence. For epitope analysis, 3'-deletional series of apo(a)VP cDNA were constructed, and expression products of them were analyzed for the binding MAb(a)20 and MAb(a)23 do. It has been revealed that distinct epitopes were recognized by the two MAbs: MAb(a)23 (gamma2b, kappa) bound to the V region about 60 aa downstream from the N-terminal, and MAb(a)20 (gamma1, kappa) bound to the P region close to the C-terminal. A one step-sandwich ELISA system for Lp(a) was developed using MAb(a)20 as a capturing antibody and horseradish peroxidase (HRP)-coupled MAb(a)23 as a detecting antibody. The assay was found to be sensitive and useful for detecting Lp(a) in the range of 4-150 microg/dL (80 pM-3 nM).

2703 related Products with: Production and characterization of monoclonal antibodies directed to the kringle V and protease domains of human apolipoprotein(a).

Rabbit Anti-Human Androge Goat Anti-Human Androgen VEGFR-2 (Human, monoclona VEGFR-2 (Human, monoclona VEGFR-1 (Human Mouse, Mon VEGFR-1 (Human, monoclona Hsp90 total Monoclonals A Anti AGO2 Human, Monoclon Anti AGO2 Human, Monoclon Anti Human AGO3, Monoclon Viral antibodies, anti-R anti FAS IgG1 (monoclonal

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Functional characterization of monoclonal antibody inhibitors of alpha 2-antiplasmin that accelerate fibrinolysis in different animal plasmas.

In humans with acute thrombotic disease, thrombi often appear to resist fibrinolysis induced by plasminogen activators. To examine the potential role of alpha 2-antiplasmin (alpha 2AP) in thrombus resistance in vivo, we generated monoclonal antibody inhibitors of alpha 2AP. In a somatic cell fusion, 99 hybridomas were obtained that produced MAbs that bound to human 125I-alpha 2AP in a capture assay. Screening assays showed that 3 of these MAbs, 49, 70, and 77, neutralized the function of alpha 2AP. Immunoblotting experiments indicated that these MAbs recognized an epitope present in native alpha 2AP that was destroyed by denaturation with SDS. Each of these MAbs fully inhibited the binding of the other MAbs to alpha 2AP, but none of them competed with the binding of another anti-alpha 2AP MAb, RWR. When tested for their binding to nonhuman alpha 2APs in plasmas, all three MAbs were strongly crossreactive with all primate plasmas tested but showed an idiosyncratic pattern of binding to alpha 2AP in other plasmas, suggesting unique fine epitope specificities. In human plasma, all three MAbs amplified the lysis of human plasma clots induced by plasminogen activators, increasing the potency of urokinase by nearly 50- to 100-fold. These MAbs also markedly amplified the lysis of clots from baboon, cynomolgus, african green monkey plasmas, and to a lesser extent, ferret and dog. By virtue of their ability to potently inhibit alpha 2AP in other animal plasmas, these MAbs should be useful for examining the role of alpha 2AP in thrombus resistance to fibrinolysis in vivo.

2281 related Products with: Functional characterization of monoclonal antibody inhibitors of alpha 2-antiplasmin that accelerate fibrinolysis in different animal plasmas.

Monoclonal anti-human CD5 Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon HIV1 integrase antibody, alpha 1 Antichymotrypsin alpha 1 Antichymotrypsin alpha 1 Antitrypsin antib hCG alpha antibody, Monoc hCG alpha antibody, Monoc FSH alpha antibody, Monoc

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Screening panels of monoclonal antibodies using phage-displayed antigen.

A procedure is described to screen panels of hybridomas or purified monoclonal antibodies using antigen displayed on the surface of filamentous bacteriophage. In this system, samples containing murine monoclonal antibodies are incubated with phage-displayed antigen in microtiter plates coated with rabbit anti-mouse IgG, and bound antibody-phage complex is detected with horseradish peroxidase-sheep anti-phage M13 conjugate. The assay has been validated with a panel of 16 monoclonal antibodies directed against human plasminogen, using phage-displayed miniplasmin-(ogen) (amino acids Ala444 through Asn791 comprising kringle 5 and the proteinase domain of plasminogen) or microplasminogen (amino acids Ala543 through Asn791 comprising the proteinase domain). Six monoclonal antibodies were identified directed against miniplasminogen and miniplasmin; this was confirmed using a microtiter plate coated with antigens. One of these monoclonal antibodies (MA-42B12) did not react with microplasminogen, suggesting that its epitope is comprised within the kringle 5 domain. This test is rapid and sensitive (detecting 10-20 ng/ml of monoclonal antibody), and screening can be performed using phage-displayed zymogens or active enzymes or selected domains thereof. The procedure eliminates the need for large amounts of purified antigen for screening. Furthermore, immunization can be performed with partially purified antigen because only antibodies raised against the antigen of interest will be identified with the use of phage-displayed antigen. Therefore, this test may offer distinct advantages over the classical one-site enzyme-linked immunosorbent assay using antigen-coated microtiter plates.

2814 related Products with: Screening panels of monoclonal antibodies using phage-displayed antigen.

Viral antibodies: anti-H Rat monoclonal anti mouse Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti Ago1, Monoclonal Ant Anti PIWIL1, Monoclonal A Anti AGO2 Mouse, Monoclon Anti Ago1, Monoclonal Ant Anti Human AGO3, Monoclon Viral antibodies, anti-R anti HBsAg surface antige

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Urokinase-type plasminogen activation in three human breast cancer cell lines correlates with their in vitro invasiveness.

In order to invade and spread cancer cells must degrade extracellular matrix proteins. This degradation is catalysed by the concerted action of several enzymes, including the serine protease plasmin. Several experimental studies have shown that inhibition of plasmin formation reduces cancer cell invasion and metastasis, indicating a critical role of this proteolytic pathway in these processes. In order to further study the role of plasmin in cancer progression, we have characterized urokinase-type plasminogen activator (uPA) mediated plasmin formation in three human breast cancer cell lines. Using monoclonal antibodies against uPA and its receptor uPAR, we have investigated the contribution of uPA and uPAR to invasive capacity in an in vitro invasion assay. MDA-MB-231 BAG cells were found to express high protein levels of uPA, uPAR and PAI-1. MDA-MB 435 BAG cells produced low amounts of uPA, PAI-1 and moderate amounts of uPAR, whereas MCF-7 BAG cells showed low levels of uPA, uPAR and PAI-1 protein. In a plasmin generation assay MDA-MB-231 BAG cells were highly active in mediating plasmin formation, which could be abolished by adding either an anticatalytic monoclonal antibody to uPA (clone 5) or an anti-uPAR monoclonal antibody (clone R3), which blocks binding of uPA to uPAR. The two other cell lines lacked the capacity to mediate plasmin formation. In the Matrigel invasion assay the cells showed activity in this order: MCF-7 BAG < MDA-MB-435 BAG < MDA-MB-231 BAG. Testing MDA-MB-231 BAG cells in the Matrigel invasion assay revealed that invasion could be inhibited in a dose-dependent manner either by the clone 5 uPA antibody or by the clone R3 uPAR antibody, suggesting that the cell surface uPA system is actively involved in this invasive process. It is concluded that these three cell lines constitute a valuable model system for in vitro studies of the role of cell surface uPA in cancer cell invasion and has application in the search for novel compounds which inhibit mechanisms involved in uPA-mediated plasmin generation on cancer cells.

1076 related Products with: Urokinase-type plasminogen activation in three human breast cancer cell lines correlates with their in vitro invasiveness.

High density (188 cases 2 High density (188 cases 2 Human breast invasive duc Top 4 types of cancer (co Top 4 types of cancer (co CELLKINES Natural Human I Interferon-a Receptor Typ Macrophage Colony Stimula Macrophage Colony Stimula Cultrex In Vitro Angiogen Human integrin aVb3, affi Breast cancer tissue arra

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Establishment and characterization of permanent human endothelial cell clones.

Though vasculitic diseases have been claimed to be associated with anti-endothelial cells antibodies (AECA), there is a widespread awareness of the limitations of the tests currently in use. Our objective was therefore to establish clones, in the hope that some of them would express disease-specific membrane autoantigens. Two EC lines and 7 clones were established by fusing human umbilical vein EC with epithelial A549/8 cells, and cloning by limiting dilution. An additional clone was derived from the EA.hy 926 cell line. All clones carried EC markers, such as thrombomoduline (TM) and platelet-EC adhesion molecule 1 but differed from each other, depending on whether they expressed HLA class II antigen, LFA-1, thrombospondin receptor or von Willebrand factor (vWf) antigen. Clones were also characterized by their ability to release tissue plasminogen activator, interleukin 6, TM and vWf. This panel is meant to distinguish reactivities of AECA.

2431 related Products with: Establishment and characterization of permanent human endothelial cell clones.

Human Umbilical Vein Endo GFP Expressing Human Umbi Mitochondria GFP Tag Huma Plasma Membrane GFP Tag H RFP Expressing Human Umbi Human Brain Microvascular GFP Expressing Human Brai Human Dermal Lymphatic Mi GFP Expressing Human Derm Human Glomerular Microvas GFP Expressing Human Glom RFP Expressing Human Glom

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Preparation, characterization and application of monoclonal antibodies against PAI-1.

Six hybridoma cell lines (AP1-AP6) secreting monoclonal antibodies (McAb) against PAI-1 were obtained by fusing the murine myeloma cell line SP2/0 with the spleen cells from Balb/c mouse immunized with recombinant PAI-1 expressed in E. coli. These antibodies were purified by SPA affinity chromatography. All McAbs recognized rPAI-1 and PAI-1 from the human hepatoma cell line HepG2. The titers of ascites were more than 10(6). The antibody-antigen affinity constants (Kaff) for anti-PAI-1 McAb measured by EIA were between 3.45 x 10(7)-1.05 x 10(10) M. AP2 and AP3 McAbs were effective in quenching the activity of PAI-1. Partial quenching of PAI-1 activity was achieved with AP4, AP5 and AP6 McAbs respectively. AP1 McAb had no effect upon PAI-1 activity. Three of the six McAbs (AP1, AP4 and AP5) bound to the PAI-1/t-PA complex, while the others did not. The PAI-1 was purified 51 folds to homogeneity from serum free medium of HepG2 with the recovery rate of 92% by one-step procedure using Sepharose 4B conjugated with anti-PAI-1 McAb (AP1, AP3 and AP4). A sandwich ELISA for the measurement of PAI-1 antigen in human plasma was developed, based on anti-PAI-1 McAb against non-overlapping epitopes. The mean value of plasma PAI-1 for the healthy donors was 24.7 +/- 7.75 ng/ml measured by ELISA.

2571 related Products with: Preparation, characterization and application of monoclonal antibodies against PAI-1.

MOUSE ANTI BOVINE ROTAVIR Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti Ago1, Monoclonal Ant Anti PIWIL1, Monoclonal A Anti AGO2 Mouse, Monoclon Anti Ago1, Monoclonal Ant Anti Human AGO3, Monoclon Viral antibodies, anti-R Signal Transduction Anti Signal Transduction Anti

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A new culture system for the cultivation of mammalian cells for the production of several biologically active substances.

We recently developed a new dialysis culture system (termed LIFROC-device) for the cultivation of lymphokine-activated killer cells (Murata et al., 1990, 1991). In the present study, we applied the LIFROC-device (400 ml culture vessel) to the cultivation of mammalian cells for the production of biologically active substances. We cultured mouse-mouse hybridoma TP-709, secreting anti-tisse plasminogen activator (tPA) monoclonal antibody (mAb), recombinant CHO GT19, secreting hGH, and human melanoma Bowes cells, secreting tPA. With the LIFROC-device, TP-709 grew to a maximal cell density of 3.8 x 10(6) cells/ml and and produced 480 micrograms/ml (192 mg in total) of mAb. GT19 reached a cell density of 2.2 x 10(6) cells/ml and produced 302 micrograms/ml (120 mg in total) of hGH. Bowes cells expanded to 4.4 x 10(6) cells/ml and secreted 8.5 micrograms/ml (3.3 mg in total) of tPA. The protein concentration in the culture broths of the LIFROC-device became 7-200 times higher than that of batch culture. Thus, the LIFROC-device can be applied to protein production as well as cell growth with high efficiency.

1562 related Products with: A new culture system for the cultivation of mammalian cells for the production of several biologically active substances.

Glycosylated Human PAI-1 Mouse PAI-1 (wild type ac Mouse PAI-1 (wild type ac Porcine PAI-1 (wild type Porcine PAI-1 (wild type Rabbit PAI-1 (wild type a Rabbit PAI-1 (wild type a Rat PAI-1 (wild type acti Rat PAI-1 (wild type acti AccuPrep Genomic DNA Extr MOUSE ANTI BOVINE ROTAVIR Bone Morphogenetic Protei

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